I have human brain tissue samples that I lyse with a nuclear extraction buffer and then the pellet I solubilized in 8M urea. I run samples in Tris-Glycine buffer in premade 4-12% gels. Ran the gel at 80V. I use iBlot2 to transfer, P0 setting. Then, I did total protein stain with Revert total protein stain.

When I transfer, I see that the lanes (total protein) are not straight. What is causing them to not transfer straight?

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